CN115152702A - Bee acute oral toxicity test and acute contact toxicity test method - Google Patents
Bee acute oral toxicity test and acute contact toxicity test method Download PDFInfo
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- CN115152702A CN115152702A CN202210716618.2A CN202210716618A CN115152702A CN 115152702 A CN115152702 A CN 115152702A CN 202210716618 A CN202210716618 A CN 202210716618A CN 115152702 A CN115152702 A CN 115152702A
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- 238000012360 testing method Methods 0.000 title claims abstract description 39
- 230000001154 acute effect Effects 0.000 title claims abstract description 31
- 231100000820 toxicity test Toxicity 0.000 title claims abstract description 26
- 231100000460 acute oral toxicity Toxicity 0.000 title claims abstract description 20
- 238000010998 test method Methods 0.000 title claims description 14
- 239000007788 liquid Substances 0.000 claims abstract description 110
- 241000257303 Hymenoptera Species 0.000 claims abstract description 63
- 239000003814 drug Substances 0.000 claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 16
- 235000003642 hunger Nutrition 0.000 claims abstract description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 12
- 229930006000 Sucrose Natural products 0.000 claims description 12
- 239000005720 sucrose Substances 0.000 claims description 11
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 10
- 230000000384 rearing effect Effects 0.000 claims description 8
- 230000037351 starvation Effects 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 5
- 239000001569 carbon dioxide Substances 0.000 claims description 5
- 206010002091 Anaesthesia Diseases 0.000 claims description 4
- 230000037005 anaesthesia Effects 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 230000006378 damage Effects 0.000 abstract description 5
- 231100000419 toxicity Toxicity 0.000 abstract description 3
- 230000001988 toxicity Effects 0.000 abstract description 3
- 230000000903 blocking effect Effects 0.000 description 54
- 241000256844 Apis mellifera Species 0.000 description 26
- 239000000243 solution Substances 0.000 description 11
- 229960004793 sucrose Drugs 0.000 description 10
- 239000012085 test solution Substances 0.000 description 6
- 235000005911 diet Nutrition 0.000 description 5
- 230000004308 accommodation Effects 0.000 description 4
- 230000037213 diet Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 2
- 206010058667 Oral toxicity Diseases 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 231100000215 acute (single dose) toxicity testing Toxicity 0.000 description 1
- 238000011047 acute toxicity test Methods 0.000 description 1
- 238000001949 anaesthesia Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000004247 hand Anatomy 0.000 description 1
- 239000011344 liquid material Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 231100000418 oral toxicity Toxicity 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000012502 risk assessment Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 210000003813 thumb Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K53/00—Feeding or drinking appliances for bees
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Infusion, Injection, And Reservoir Apparatuses (AREA)
Abstract
The invention discloses a method for testing acute oral toxicity and acute contact toxicity of bees, which comprises the steps of collecting bees, conducting hunger treatment on the collected bees, directly conducting the acute oral toxicity and acute contact toxicity tests on the hunger-treated bees, feeding the bees by a feeder in the experimental process, avoiding damages to the bees, opening an acute radial toxicity pattern of the bees without anaesthetizing the bees, solving the problem of incomplete liquid medicine taking by the feeder, and avoiding liquid medicine leakage.
Description
Technical Field
The invention relates to the technical field of bee experiment methods, in particular to a bee acute oral toxicity test method and an acute contact toxicity test method.
Background
Acute oral toxicity test of bees: dispersing the tested substances with different dosages in sucrose solution for feeding adult worker bees, measuring the consumption of the liquid medicine, and feeding the sucrose solution without the tested substances after the liquid medicine is consumed.
Acute contact toxicity test of bees: after bees are narcotized, test liquid medicines with different concentrations are dripped on the middle chest back plate of the bees for test, after the solvent is volatilized, the bees are transferred into a test cage, and the test cage is soaked with appropriate amount of cane sugar water for feeding.
Traditional bee acute toxicity test bee collection requires whole spleen anesthesia. Then every 10 bees are transferred to 1 bee cage, and the revived bees need to be anaesthetized for two times or more times, which can cause great harm to the bees. The experimental centrifuging tube or syringe of using of oral toxicity of honeybee acute, the centrifuging tube needs the people to prick the hole, and the syringe needs the certain time people to go to push away and makes the liquid medicine hang and can not fall, when weighing the feeding tube and when infecting the poisonous honeybee, all has the risk of liquid medicine drip, has the honeybee simultaneously and can't get the liquid medicine and eat clean drawback, and then influences experimental accuracy, and the pesticide produces very big influence to the risk assessment of honeybee.
Disclosure of Invention
In order to solve the technical problems in the background technology, the invention provides a method for acute oral toxicity test and acute contact toxicity test of bees.
The invention provides a method for acute oral toxicity test and acute contact toxicity test of bees, which comprises the following steps:
starvation treatment:
placing a certain amount of bees into a rearing cage, and carrying out starvation treatment for 1-3 h under the dark condition;
carrying out bee acute oral toxicity test:
placing the starved bees into a feeding box;
weighing the feeder filled with the experimental liquid medicine in the feeder of the experimental liquid medicine, putting the weighed feeder into a feeding box, taking out the feeder after 3-6 hours, and weighing the feeder;
after the bees in the feeder suck a certain amount of the experimental liquid medicine, placing the feeder filled with the sucrose solution in a feeding box;
performing an acute contact toxicity test on bees:
placing the hungry bees into a centrifugal tube, introducing carbon dioxide into the centrifugal tube for anesthesia for 3-5 s, placing the bees on a test bed, clamping wings of the bees by using tweezers, dripping 1 mu L of test liquid medicine to a mesothorax backplate of the bees by using a micro-drip apparatus or a pipettor, and transferring the test liquid medicine into a bee box after the solvent is volatilized;
a feeder filled with sucrose solution is placed in the feeding box.
The feeder can be the container that can hold the liquid material among the prior art and have the opening and be convenient for honeybee diet, and is specific, the feeder is including holding the post, it has the holding tank to hold the top of post and open, and the opening of holding tank is held the top of post and is opened the holding tank of lateral part relatively and be convenient for honeybee diet feeder liquid.
Preferably, the holding tank is cylindrical, the aperture of the holding tank is 7mm, the volume of the holding tank is slightly larger than 200 mu L, and the opening of the holding tank only allows the body of one bee to extend to the liquid in the diet holding tank in the holding tank, so that two or more bees are prevented from eating the liquid in the holding tank at the same time, and the feeder is prevented from toppling over.
Preferably, still include the panel, it has the accommodation hole to open on the panel, holds the vertical slidable mounting of post in the accommodation hole, avoids holding the post and empting when feeding the box outside, and is specific, and the panel that has certain thickness that the accommodation hole can be opened for among the prior art to the panel, avoids holding the post and empties.
Preferably, the liquid blocking device further comprises a liquid blocking ring, wherein the liquid blocking ring is mounted on the containing column and used for preventing the solution from splashing out of the containing tank, and particularly, the liquid blocking ring can be made of plastic or metal or paper material.
Preferably, the liquid blocking ring is conical, the inner diameter of the liquid blocking ring is gradually reduced from one end close to the accommodating tank to one end far away from the accommodating tank, the diameter of one end, far away from the accommodating tank, of the liquid blocking ring is 1-4mm, namely the opening aperture of the liquid blocking ring is 1-4mm, the situation that liquid in the accommodating tank is prevented from leaking from a feeder in the process that the feeder is placed in a feeding box is avoided, liquid on the inner wall of the liquid blocking ring is conveniently discharged into the accommodating tank after the feeder is placed by the conical liquid blocking ring, the diameter of one end, far away from the accommodating tank, of the liquid blocking ring is 1-4mm, and liquid is prevented from being discharged from an opening of the liquid blocking ring under the action of surface tension of the liquid.
In order to facilitate the detachment of the liquid blocking ring, preferably, the liquid blocking ring is made of thin paper, the outer side wall of the liquid blocking ring is provided with an annular scratch along the circumferential direction of the liquid blocking ring, and after the liquid blocking ring and a feeder are placed in the feeding box together, the liquid blocking ring is broken from the annular scratch by pulling the liquid blocking ring, so that the part of the liquid blocking ring separated from the feeder is taken out of the feeding box.
Preferably, it includes inner ring and outer loop to keep off the liquid ring, the inner ring with the holding tank intercommunication, the outer loop suit is fixed the outer wall of inner ring, the outer loop is fixed hold the outer wall of post, annular mar is seted up on the outer loop, just annular mar with the up end that holds the post is in the coplanar, and it is connected and is convenient for simultaneously keep off the liquid ring and be convenient for keep off the liquid ring along annular mar punishment separation and be convenient for seal from holding tank opening part diet liquid with the feeder to be convenient for.
Preferably, the lower extreme that holds the post is connected through the bottom plate of screw thread mode with feeding the box, prevents on the one hand that the feeder from empting, and on the other hand is convenient for keep off the dismantlement of liquid ring, and is convenient for keep off the liquid of liquid ring inner wall and arranges the increase in the holding tank and detect the precision.
Preferably, the bees in the rearing cage in the step S1 are worker bees.
Preferably, be fixed with pneumatic quick-operation joint on the lid of centrifuging tube in step S5, let in carbon dioxide in the centrifuging tube through the pneumatic quick-operation structure of hose connection, be convenient for improve work efficiency with current artificial pricking hole to the centrifuging tube relatively.
Specifically, in step S3, the test solution is a mixture of sucrose and the test substance.
According to the method for the bee acute oral toxicity test and the acute contact toxicity test, disclosed by the invention, the quick collection of bees can be completed, the damage to the bees is avoided, the acute radial toxicity pattern of the bees can be opened without anaesthetizing the bees, the problem of incomplete liquid medicine taking is solved by using the feeder, and the liquid medicine is prevented from dripping and leaking.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
FIG. 1 is a schematic structural diagram of a centrifuge tube in the method;
FIG. 2 is a schematic view of the connection structure of the display plate and the feeder of the present invention;
FIG. 3 is a schematic structural view of the liquid-blocking ring and the receiving column after they are connected according to the present invention;
FIG. 4 is a schematic diagram of the structure of the split feeding box of the invention after the front plate and the rear feeder are placed in the feeding box;
FIG. 5 is a schematic structural view of a feeder and a feeding box after a broken liquid blocking ring is taken out of a front plate of the feeding box cut open according to the present invention.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar designations denote like or similar elements or elements having like or similar functionality throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
It will be understood that the terms "central," "longitudinal," "transverse," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in an orientation or positional relationship indicated in the drawings for convenience and simplicity of description only and are not intended to indicate or imply that the device or element so referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be considered as limiting the invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; may be mechanically coupled, may be electrically coupled or may be in communication with each other; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
In the present invention, unless expressly stated or limited otherwise, the first feature "on" or "under" the second feature may be directly contacting the second feature or the first and second features may be indirectly contacting each other through intervening media. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
A bee acute oral toxicity test and acute contact toxicity test method comprises the following steps:
starvation treatment:
s1: putting on the protective clothing, taking out the honeycomb from the beehive, shaking into a bee rearing cage according to the required number of bees after no queen bee exists, quickly closing the zipper of the rearing cage, and carrying out starvation treatment for 2h in dark condition;
s2, carrying gloves, extending hands into the rearing cage through sleeves of the bee rearing cage, using a 50mL centrifugal tube 3, enabling a tube opening to be tightly attached to the upper part of the rearing cage, buckling the bees into the centrifugal tube 3, enabling the bees to enter the bottom of the centrifugal tube 3 by slightly shaking the centrifugal tube 3, taking 10 bees, and then pressing the tube opening with a thumb to prevent the bees from escaping;
carrying out bee acute oral toxicity test:
s300: slightly shaking the centrifugal tube 3 in the S2 to enable bees to enter the bottom of the centrifugal tube 3, aligning the opening of the centrifugal tube 3 with the opening of the feeding box 1, shaking the bees into the feeding box 1, and placing the bees subjected to hunger treatment into the feeding box 1;
s301: as shown in fig. 2, a feeder is placed on a display board 2 and numbered, the weight of the empty feeder is weighed by a ten-thousandth balance, 200 mul of test liquid medicine is transferred into the feeder by a pipette, an outer ring 51 of a liquid blocking ring 5 is fixed on the outer wall of a containing column 4 in an adhering mode, an inner ring 50 covers an opening of the containing tank 40 (as shown in fig. 3), the total weight of the feeder is weighed again (the total weight of the feeder comprises the total weight of the feeder, the liquid blocking ring 5 and the test liquid medicine), the feeder and the liquid blocking ring 5 are quickly placed into a feeding box 1 (as shown in fig. 4) by using tweezers, rong Nazhu is rotated to ensure that the lower part of the containing column 4 is in threaded connection with the bottom plate of the feeding box 1, the top of the liquid blocking ring 5 is pulled by the tweezers to ensure that the liquid blocking ring 5 is broken from a scratch 510 of the annular, the broken liquid blocking ring 5 is partially taken out (as shown in fig. 5), the test liquid medicine taken out by bees freely from the containing tank 40, the feeding device after 4h, the weighing process, the test liquid medicine taken out and the test liquid medicine taken out in the test solution of the honey bee is weighed acute test (the test solution) in the test solution and the test solution in the conventional test solution step);
s302: after the feeder filled with the test liquid medicine is taken out, the feeder filled with the sucrose solution is placed in the feeding box 1 through tweezers, the same as the steps, the sucrose solution is placed in the feeder, then the notch of the holding tank 40 of the feeder is coated through the liquid blocking ring 5, after the feeder is fixed in the feeding box 1, the liquid blocking ring 5 is pulled to break from the annular scratch 510, and the broken liquid blocking ring 5 is taken out; the later experimental process is the same as the existing bee acute oral toxicity experimental process, and the vital signs of the bees are observed and recorded;
performing an acute contact toxicity test on bees:
s310, covering a cover on the centrifugal tube 3 in the S2, arranging a pneumatic quick connector (shown in figure 1) on the cover of the centrifugal tube 3, connecting one end of the pneumatic quick connector with a carbon dioxide gas cylinder through a hose, extending the other end of the pneumatic quick connector into an opening of the cover of the centrifugal tube 3, unscrewing a main valve of a steel cylinder, adjusting a pressure valve to 0.02Mpa, placing bees on a test table after anaesthetizing for 4S, clamping wings of the bees by using tweezers, dripping 1 mu L of test liquid medicine to a mesothorax plate of the bees by using a micro-dropping instrument or a pipettor, and transferring the test liquid medicine to the feeding box 1 after the solvent is volatilized;
s311, placing a feeder filled with a sucrose solution in the feeding box 1; the same as the above steps, the holding tank 40 notch of the feeder is coated by the liquid blocking ring 5 after the sucrose solution is always placed in the feeder, and after the feeder is fixed in the feeding box 1, the liquid blocking ring 5 is broken from the annular scratch 510 by pulling and the broken liquid blocking ring 5 is taken out.
As shown in fig. 2-5, the feeder in the above steps of the method comprises a holding column 4, the top end of the holding column 4 is provided with a holding tank 40, the opening of the holding tank 40 is arranged at the top of the holding column 4 opposite to the holding tank 40 at the side part of the holding tank 40, so that the bees can eat the liquid in the feeder, the holding tank 40 is cylindrical, the aperture of the holding tank 40 is 7mm, the volume of the holding tank 40 is slightly larger than 200 μ L, and the opening of the holding tank 40 only allows the body of one bee to extend into the liquid in the eating holding tank 40 in the holding tank 40, so as to prevent two or more bees from eating the liquid in the holding tank 40 at the same time, and prevent the feeder from falling;
in order to further avoid the tilting of the feeder, in this embodiment, it is preferable that the lower end of the containing column 4 is provided with an external thread, and the bottom plate of the feeding box 1 is provided with a threaded hole which is matched with the external thread at the lower end of the containing column 4;
the feeding device is characterized by further comprising a display plate 2, wherein the display plate 2 is provided with a containing hole 20, the containing column 4 is vertically and slidably arranged in the containing hole 20, the containing column 4 is prevented from toppling over when being outside the feeding box 1, and when the feeder is not placed in the feeding box, the feeder is placed in the containing hole 20 of the display plate 2;
the feeding box also comprises a liquid blocking ring 5, wherein the liquid blocking ring 5 is arranged on the containing column 4 and used for preventing a solution from splashing out of the containing tank 40, the liquid blocking ring 5 covers an opening of the containing tank 40, the liquid blocking ring 5 is conical, the inner diameter of the liquid blocking ring 5 is gradually reduced from one end close to the containing tank 40 to one end far away from the containing tank 40, the diameter of one end, far away from the containing tank 40, of the liquid blocking ring 5 is 1-4mm, namely the aperture of the opening of the liquid blocking ring 5 is 1-4mm, so that the situation that the liquid in the containing tank 40 is prevented from leaking out of the feeder in the process that the feeder is placed in the feeding box 1 is avoided, the conical liquid blocking ring 5 is convenient for discharging the liquid on the inner wall of the liquid blocking ring 5 into the containing tank 40 after the feeder is placed, and the diameter of one end, far away from the containing tank 40, of the liquid blocking ring 5 is 1-4mm, so that the liquid is prevented from being discharged out of the opening of the liquid blocking ring 5 under the action of the surface tension of the liquid;
the liquid blocking ring 5 is made of thin paper, the outer side wall of the liquid blocking ring 5 is provided with an annular scratch 510 along the circumferential direction of the liquid blocking ring, and when the liquid blocking ring 5 and a feeder are placed in the feeding box 1 together, the liquid blocking ring 5 is broken from the annular scratch 510 by pulling the liquid blocking ring 5, so that the part of the liquid blocking ring 5 separated from the feeder is taken out of the feeding box 1;
the liquid blocking ring 5 comprises an inner ring 50 and an outer ring 51, the inner ring 50 is communicated with the accommodating groove 40, the lower end of the inner ring 50 is abutted against the upper end face of the accommodating column, the inner ring 50 is conical, the outer ring is cylindrical, the outer ring 51 is sleeved and fixed on the outer wall of the lower end of the inner ring 50, the outer ring 51 is fixed on the outer wall of the accommodating column 4, the annular scratch 510 is arranged on the outer ring 51, the annular scratch 510 and the upper end face of the accommodating column 4 are in the same plane, the liquid blocking ring 5 is conveniently connected with a feeder, and meanwhile, the liquid blocking ring 5 is conveniently separated along the annular scratch 510 so as to be convenient for sealing the dietary liquid at the opening of the accommodating groove 40;
when placing the feeder, will hold post 4 and place the screw hole department of feeding box 1 and then rotate through tweezers and hold post 4, the axis that makes to hold post 4 along vertical direction rotates and makes the lower extreme that holds post 4 and the bottom plate of placing the box pass through the screw thread mode and be connected, the liquid stream that also is convenient for keep off the liquid ring 5 inner wall at the rotation in-process flows to holding tank 40 in, then the shredding keeps off liquid ring 5 and takes out the shredding keep off liquid ring 5 from feeding box 1, will hold post 4 and take out and place in the accommodation hole 20 of panel 2 through tweezers after a certain time.
The invention can rapidly collect bees and avoid the damage to the bees. The bee acute oral toxicity test can be carried out without anaesthetizing bees, and the feeder can solve the problem of incomplete liquid medicine taking and avoid the risk of liquid medicine leakage. The bee acute contact toxicity test can be used for independently anaesthetizing bees repeatedly, so that the integral anaesthesia is avoided, and the harm to the bees is reduced; the whole set of new method makes the test more scientific and reasonable.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (10)
1. A bee acute oral toxicity test and acute contact toxicity test method is characterized by comprising the following steps:
starvation treatment:
placing a certain amount of bees into a rearing cage, and carrying out starvation treatment for 1-3 h under the dark condition;
carrying out bee acute oral toxicity test:
placing the starved bees into a feeding box;
weighing the feeder filled with the experimental liquid medicine in the feeder of the experimental liquid medicine, putting the weighed feeder into a feeding box, taking out the feeder after 3-6 hours, and weighing the feeder;
after the bees in the feeder suck a certain amount of the experimental liquid medicine, placing the feeder filled with the sucrose solution in a feeding box;
carrying out an acute contact toxicity test on bees:
placing the hungry bees into a centrifugal tube, introducing carbon dioxide into the centrifugal tube for anesthesia for 3-5 s, placing the bees on a test bed, clamping wings of the bees by using tweezers, dripping 1 mu L of test liquid medicine to a mesothorax backplate of the bees by using a micro-drip apparatus or a pipettor, and transferring the test liquid medicine into a bee box after the solvent is volatilized;
a feeder filled with sucrose solution is placed in the feeding box.
2. The bee acute oral toxicity test and acute contact toxicity test method of claim 1, wherein the feeder comprises a receiving column with a holding trough at the top end.
3. The bee acute oral toxicity test and acute contact toxicity test method of claim 2, wherein the aperture of the holding tank is 7mm.
4. The bee acute peroral toxicity test and acute contact toxicity test method of claim 2, further comprising a display board, wherein the display board is provided with a containing hole, and the containing column is vertically and slidably installed in the containing hole.
5. The bee acute oral toxicity test and acute contact toxicity test method of claim 2, further comprising a liquid-blocking ring mounted on said containment column and configured to prevent the solution from splashing out of said containment tank.
6. The bee acute peroral toxicity test and acute contact toxicity test method of claim 5, wherein said liquid-blocking ring is tapered, and the inner diameter of the liquid-blocking ring becomes gradually smaller from the end near the holding tank to the end far from the holding tank.
7. The bee acute peroral toxicity test and acute contact toxicity test method of claim 5 or 6, wherein the outside wall of the liquid-barrier ring is provided with an annular scratch along its circumference.
8. The bee acute peroral toxicity test and acute contact toxicity test method of claim 7, wherein said liquid-blocking ring comprises an inner ring and an outer ring, said inner ring is connected to said holding tank, said outer ring is sleeved and fixed on the outer wall of said inner ring, said outer ring is fixed on the outer wall of said holding column, said annular scratch is opened on said outer ring, and said annular scratch is in the same plane with the upper end surface of said holding column.
9. The bee acute oral toxicity test and acute contact toxicity test method according to any of claims 2-6, wherein the lower end of said containment column is connected to the bottom plate of the feeding box by means of screw threads.
10. The method of claim 1, wherein a pneumatic quick connector is fixed on the cover of the centrifuge tube in step S5, and carbon dioxide is introduced into the centrifuge tube through a hose connected to the pneumatic quick connector.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210716618.2A CN115152702B (en) | 2022-06-23 | 2022-06-23 | Bee acute oral toxicity test and acute contact toxicity test method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210716618.2A CN115152702B (en) | 2022-06-23 | 2022-06-23 | Bee acute oral toxicity test and acute contact toxicity test method |
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| CN115152702A true CN115152702A (en) | 2022-10-11 |
| CN115152702B CN115152702B (en) | 2023-11-03 |
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| JPH0423653U (en) * | 1990-06-21 | 1992-02-26 | ||
| JPH06329160A (en) * | 1993-05-17 | 1994-11-29 | Keisuke Ito | Complex container retaining freshness after unseal and its production |
| JPH119657A (en) * | 1997-06-24 | 1999-01-19 | Eisai Co Ltd | Mouth piece for medicine bottle |
| KR19990012774U (en) * | 1997-09-12 | 1999-04-15 | 이태환 | Snap cap of the container |
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